| Object:On the basis of previous studies for hepatitis B virus (hepatitis B virus, HBV) infection in high-risk populations of liver cancer at the same time drinking, establish alcoholic HBV transgenic mice HCC precancerous lesion model, exploring new ways to establish an animal model of HCC precancerous lesions, lay the foundation for further study of the molecular mechanisms of the development of liver cancer;Early treatment for precancerous lesions of liver cancer model mice fed Gehuajiecheng decoction, observe the mouse model of alanine aminotransferase (ALT), aspartate aminotransferase(AST), glutathione S-transferase (GST), ?-glutamyl transferase (GGT) expression in serum level and for cyclin D1(CyclinDl), cyclin-dependent kinase 4(CDK4) and p27 expression.Methods:1. HCC precancerous lesions Mouse Model:The 32 HBV transgenic mice and 32 BALB/c non-transgenic mice be adaptive feeding for 5 days, HBV transgenic mice and BALB/c mice were randomly divided into ordinary 16 weeks,22 weeks in the control group,16 week,22 weeks model group (n=8). Model group was given 53% (v/v) liquor dose of 10ml/(kg · d) orally once a day;the control group received an equal volume of distilled water once a day. Adopting clean diet during feeding, a weight weighing every 5 days, intragastric dose adjustment based on changes in body weight. Mice were drawn at 16,22 weekends, mice were fasted for 24h prior to sacrifice, to take the eye blood, the mice were sacrificed by cervical dislocation, the liver of mice immediately after death, after weighing in 4% paraformaldehyde;thymus weighed to calculate the thymus index; During the experimental observation:mouse activity,mental state, fur gloss, loss of appetite, etc.2. The impact of Gehuajiecheng decoction for alcoholic HBV transgenic mice model of HCC precancerous lesions of related indicators:The 24 HBV transgenic mice adaptive feeding for 5 days, were randomly-divided into control group, model group and treatment group(n= 8). The treatment group conversion method based on animal pharmacology experiments medication, adult dosage of conventional amounts are translated experiments in mice and experimental study group preliminary results that the treatment group were 0.4g(crude drug)/10g(body mass) Gehuajiecheng decoction volume of 5ml/kg, control group and model group were given an equal volume of distilled water once a day;30min after treatment group and model group was given 53%(v/v)Double pot liquor,10ml /kg, once a day, the control group received an equal volume of distilled wate. Adopting clean diet during feeding. Mice in each group were drawn at 22 weekend, mice were fasted for 24h prior to sacrifice, to take the eye blood, the mice were sacrificed by cervical dislocation, the liver of mice immediately after death, after weighing in 4% paraformaldehyde;Thymus weighed to calculate the thymus;During the experimental observation: mouse activity, mental state, fur gloss, loss of appetite, etc. By enzyme-linked immunosorbent assay (ELISA) to detect mouse serum AST, ALT, GST and GGT; Detecting expression levels in liver tissue CyclinDl, CDK4 and P27 using immunohistochemistry.Results:(1)Alcohol on the body weight of mice before and after the experiment, liver weight and liver index, thymus weight and thymus index of influence: the model group compared with the same period of the same experimental strain of mice in the control group before and after the experiment the degree of weight gain than the control group of mice, the liver and liver index was significantly heavier than control mice, the thymus and thymus of mice was significantly lower than the control group, with statistical significance (P<0.05);The same experimental period transgenic mice model compared with normal mice model group, experiment the degree of weight gain before and after the normal mice was significantly lower than the model group, liver weight and liver index significantly increased thymus weight and thymus index was significantly lower, with statistical significance (P<0.05).22 weeks transgenic mouse models to suppress the most obvious body weight, liver weight and liver index significantly increased, thymus weight and thymus index was significantly reduced.(2) The impact Gehuajiecheng decoction of ALT, AST, GST, GGT:Compared with the model group, the control group and the treatment group indexes have significant differences (P<0.05). Model group serum ALT, AST, GST, GGT secretion compared with the control group were significantly increased (P<0.05);The treatment group compared with the model group was significantly decreased(P<0.05).(3)Gehuajiecheng decoction affect of precancerous lesions of liver-related factors:Compared with the control group, the model group CyclinDl, CDK4 expression content was significantly increased(P<0.05); Compared with the model group, the treatment group CyclinDl, CDK4 expression content was significantly lower (P<0.05). Compared with the control group, the expression of p27 content in model group was significantly lower liter (P<0.05); Compared with the model group, the treatment group P27 expression content was significantly increased (P <0.05).Conclusion:(1)Large quantities of alcohol to stimulate that the growth of mice significantly inhibited, damage the liver, increased liver weight and liver index;immune organ damage, thymus weight and thymus index was significant reduced.(2)Gehuajiecheng decoction can effectively reduce alcohol-induced liver injury in mice serum ALT, AST, GST, GGT activity increased, with hepatoprotective effect.(3) Gehuajiecheng decoction can down the expression of Cyclin Dl and CDK4, upregulation of p27 expression.Through the above experimental results, Gehuajiecheng decoction influence the mouse liver precancerous lesions related factors may be: stabilize cell membranes, protecting liver cells, improve liver function; by down-regulating the expression of oncogenes CyclinDl and CDK4, upregulate the expression of the tumor suppressor gene p27 reduction further deterioration of the liver precancerous lesions, and thus to protect and reversal. |
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